Sensitive Biomarkers For Early Diagnosis Research Articles

MicroRNA mmu-miR-511-5p: A promising Diagnostic Biomarker in Experimental Toxoplasmosis Using Different Strains and Infective Doses in Mice with Different Immune States Before and After Treatment

PurposeSearching for a novel early diagnostic biomarker for toxoplasmosis, real-time-PCR was currently used to measure the serum mmu-miR-511-5p level in male Swiss-albino mice infected with either; ME49 or RH Toxoplasma gondii (T. gondii) strains.MethodsThree mice groups were used; (GI) constituted the non-infected control group, while (GII) and (GIII) were experimentally infected with ME49 or RH strains, respectively. GII mice were orally infected using 10 or 20 ME49 cysts (ME-10 and ME-20), both were subdivided into; non-treated (ME-10-NT and ME-20-NT) and were further subdivided into; immunocompetent (ME-10-IC and ME-20-IC) [euthanized 3-days, 1, 2, 6 or 8-weeks post-infection (PI)], and immunosuppressed using two Endoxan® injections (ME-10-IS and ME-20-IS) [euthanized 6- or 8-weeks PI], and spiramycin-treated (ME-10-SP and ME-20-SP) that received daily spiramycin, for one-week before euthanasia. GIII mice individually received 2500 intraperitoneal RH strain tachyzoites, then, were subdivided into; non-treated (RH-NT) [euthanized 3 or 5-days PI], and spiramycin-treated (RH-SP) that were euthanized 5 or 10-days PI (refer to the graphical abstract).ResultsRevealed significant upregulation of mmu-miR-511-5p in GII, one-week PI, with gradually increased expression, reaching its maximum 8-weeks PI, especially in ME-20-NT group that received the higher infective dose. Immunosuppression increased the upregulation. Contrarily, treatment caused significant downregulation. GIII recorded significant upregulation 3-days PI, yet, treatment significantly decreased this expression.ConclusionSerum mmu-miR-511-5p is a sensitive biomarker for early diagnosis of ME49 and RH infection (as early as one-week and 3-days, respectively), and its expression varies according to T. gondii infective dose, duration of infection, spiramycin-treatment and host immune status.Graphical abstract

Evaluacija ekspresije, dijagnostičkog i prognostičkog značaja hsa-miR-675-5p kod oralnog karcinoma

Introduction: Oral cancer is the most common subtype of cancer in the head and neck region, with an increasing incidence worldwide. Unfortunately, no specific biomarkers are used in everyday clinical practice. Small non-coding RNA molecules, microRNA (miRNA), are considered sensitive biomarkers for early diagnosis as well as prognosis in patients with oral cancer. Especially, microRNA derived from the H19 locus are poorly investigated for their potential role in oral cancer. Aim: The aim of this study was to evaluate expression of hsa-miR675-5p in tumor and non-tumor tissues of oral cancer patients and to associate it with pathohistological features. Material and Methods: The study group consisted of 35 patients with oral cancer. Tumor and surrounding non-tumor tissues were taken from each patient. Relative expression was measured using the quantitative reverse transcription - real time PCR method. Results: The relative expression of hsa-miR-675-5p was lower in oral cancer tumor than in non-tumor tissue suggesting its tumor suppressive role. hsa-miR-675-5p has diagnostic potential for sensitive distinction of tumor and non-tumor tissues in oral cancer patients. There was no difference in overall survival rates between patients with low and high hsa-miR-675-5pexpression, confirming that hsa-miR-675-5p cannot be used as a prognostic biomarker in patients with oral cancer. Conclusion: hsa-miR-675-5p can be considered as a potential diagnostic but not a prognostic molecular biomarker in oral cancer.

Development of lectin-based lateral flow assay for fucosylated alpha-fetoprotein.

Fucosylated alpha-fetoprotein (AFP-L3) is a more specific and sensitive biomarker for early diagnosis of hepatocellular carcinoma (HCC) than only the alpha-fetoprotein (AFP) level. Rapid and simple detection of AFP-L3 level greatly facilitates the early detection as well as the treatment of HCC, resulting in the reduction of mortality. Here, we developed a rapid and sensitive lateral flow assay (LFA) using lectin Lens culinaris agglutinin (LCA), which has a specific affinity to AFP-L3 fraction of AFP, as a biorecognition element for determination of the fucosylation of AFP. The assay is based on a sandwich format performed on a lateral flow test strip. LCA was immobilized on the membrane as a test line (T). Quantitative detection of AFP-L3 was achieved by measuring the green color intensity of captured gold nanoparticle conjugates on the T and control line (C) utilizing an in-house test strip reader. The calculated absorbance obtained by the green color intensity signals proportionally increased with AFP concentrations. The developed lectin-based LFA provided a detection limit of 0.8 ng/mL for AFP with a linear range between 1.5 and 160.0 ng/mL within an assay time of 10 min. Recoveries between 74.5% and 113.2% with relative standard deviations of 5.2%-8.7% for measuring spiked human serum were also achieved. The results reveal that the proposed assay offers a rapid, sensitive, and specific method, which is useful for development in point-of-care testing for early detection and treatment of HCC.

Circular SERPINA3 and its target microRNA-944 as potential biomarkers in hepatitis C virus-induced hepatocellular carcinoma in Egyptian population.

The most prevalent cancer in Egypt is hepatocellular carcinoma (HCC) mainly due to the infection with the hepatitis C virus. So it is critical to find sensitive biomarkers for early diagnosis of HCC and avoid post-operation tumor recurrence. Therefore, this research was designed to demonstrate the circSERPINA3 role in the regulation of microRNA-944 gene expression in HCV-related HCC cases and compare these results with circSERPINA3 and microRNA-944 gene expression levels in HCV-infected patients. Study participants were divided into three groups: healthy controls, HCV- infected, and HCV-induced HCC patients. The gene expression levels of circSERPINA3 and microRNA-944 were evaluated using Real-Time qPCR. Then the immunoblotting procedure was applied to measure the serum levels of MDM2 and E-cadherin besides, the serum concentration levels of glypican-3 and alpha-fetoprotein were measured by sandwich ELISA. The gene expression level of circSERPINA3 was significantly upregulated in both HCV-infected and HCC patients causing suppression of the antitumor effect of miR-944 and showing a lower 1-year survival rate than the participants who had low circSERPINA3 gene expression levels. Subsequently, the miR-944 downstream protein, MDM2 was remarkably upregulated, exaggerating the metastasis and oxidative stress in HCC cases. Additionally, the results confirmed the downregulation of microRNA-944 improved the progression of viral hepatitis C cases to hepatocarcinogenesis through the significantly increased serum level of the metastatic marker, E-cadherin. Although alpha-fetoprotein is a common diagnostic marker used in the diagnosis of HCC, our results showed that glypican-3 had greater sensitivity and specificity and positively correlated to the IGF-1 signaling pathway of HCC cases. Moreover, the gene expression levels of circSERPINA3 and E-cadherin in both the HCV and HCV-induced HCC were significantly positively correlated. circSERPINA3 and miR-944 were sensitive molecular markers for early diagnosis of HCC and could be prospective treatment targets for HCV-infected patients to avoid tumor recurrence in HCC cases.

Chasing the Role of miRNAs in RCC: From Free-Circulating to Extracellular-Vesicle-Derived Biomarkers.

Renal cell carcinoma (RCC) is the second most common cancer of the urinary system. The current therapeutic strategies are based on partial or total nephrectomy and/or targeted therapies based on immune checkpoint inhibitors to which patients are often refractory. Preventive and screening strategies do not exist and the few available biomarkers for RCC are characterized by a lack of sensitivity, outlining the need for novel noninvasive and sensitive biomarkers for early diagnosis and better disease monitoring. Blood liquid biopsy (LB) is a non- or minimally invasive procedure for a more representative view of tumor heterogeneity than a tissue biopsy, potentially allowing the real-time monitoring of cancer evolution. Growing interest is focused on the extracellular vesicles (EVs) secreted by either healthy or tumoral cells and recovered in a variety of biological matrices, blood included. EVs are involved in cell-to-cell crosstalk transferring their mRNAs, microRNAs (miRNAs), and protein content. In particular, transferred miRNAs may regulate tumorigenesis and proliferation also impacting resistance to apoptosis, thus representing potential useful biomarkers. Here, we present the latest efforts in the identification of circulating miRNAs in blood samples, focusing on the potential use of EV-derived miRNAs as RCC diagnostic and prognostic markers.

A glimpse of recent advances in the research of acute kidney injury

AbstractAcute kidney injury (AKI) is a major renal disease that affects 13 million people and is associated with almost 1.7 million deaths each year around the world. Previously referred to as acute renal failure, AKI is defined not only by a rapid functional decline of the kidney but also by pathological damages in kidney tissue. This updated definition contributes to modern renal medicine in at least two ways. First, the definition of AKI includes an early, preventable and treatable stage of the disease, which implies an urgent need for sensitive biomarkers for early diagnosis and treatment. Second, the AKI definition is proposed with clear criteria for diagnosis and severity grading, which are beneficial for clinical studies and the meta‐analysis of different datasets.

Molecular Mechanisms and Animal Models of HBV-Related Hepatocellular Carcinoma: With Emphasis on Metastatic Tumor Antigen 1.

Hepatocellular carcinoma (HCC) is an important cause of cancer death worldwide, and hepatitis B virus (HBV) infection is a major etiology, particularly in the Asia-Pacific region. Lack of sensitive biomarkers for early diagnosis of HCC and lack of effective therapeutics for patients with advanced HCC are the main reasons for high HCC mortality; these clinical needs are linked to the molecular heterogeneity of hepatocarcinogenesis. Animal models are the basis of preclinical and translational research in HBV-related HCC (HBV-HCC). Recent advances in methodology have allowed the development of several animal models to address various aspects of chronic liver disease, including HCC, which HBV causes in humans. Currently, multiple HBV-HCC animal models, including conventional, hydrodynamics-transfection-based, viral vector-mediated transgenic, and xenograft mice models, as well as the hepadnavirus-infected tree shrew and woodchuck models, are available. This review provides an overview of molecular mechanisms and animal models of HBV-HCC. Additionally, the metastatic tumor antigen 1 (MTA1), a cancer-promoting molecule, was introduced as an example to address the importance of a suitable animal model for studying HBV-related hepatocarcinogenesis.

Comparative Analysis between Urinary Calprotectin and Serum Creatinine for Early Detection of Intrinsic Acute Kidney Injury.

Background:Acute kidney injury (AKI) is a common and important clinical condition that may lead to chronic kidney disease if it is not diagnosed and treated in its early stages. Urinary calprotectin is a valuable recognized biomarker that can be used to differentiate prerenal and intrinsic AKI. However, till date only a few reports on urine calprotectin measurement in early diagnosis of intrinsic AKI are available. In this study, we compared the sensitivity and specificity of urinary calprotectin with those of serum creatinine in detecting early intrinsic AKI.Methods:Over 6 months period (April to October 2018), 81 of 408 patients admitted to the pediatric intensive care unit met the criteria of this cross-sectional study. Their serum creatinine and urinary calprotectin were measured on the first and third day of admission using Jaffe and Elisa radioimmunoassay methods, respectively. The AKI was defined according to the pRIFLE criteria.Results:Of the total 81 patients, 67 had the criteria of intrinsic AKI. Of these 62% were female and 38% were male. The mean age of the patients was 22 months. According to data analysis, the area under the curve of ROC of urinary calprotectin on day-1 to detect renal failure is 0.93 with the best cutoff point obtained at 530 ng/mL. The sensitivity, specificity, positive, and negative predictive values of urinary calprotectin levels in diagnosing AKI at this cutoff point are 92.5%, 92.8%, 98.4, and 72.2%, respectively. Besides, urinary calprotectin changes occur much earlier than the rising of serum creatinine.Conclusion:Urinary level of calprotectin is a very sensitive biomarker for early diagnosis of intrinsic AKI in children and it can be used in intensive care units or anywhere critically ill children admitted to detect intrinsic AKI. Besides, this study shows that urine calprotectin may be a more sensitive and specific biomarker than serum creatinine in the early phases of intrinsic AKI.

Assessment of neutrophil gelatinase-associated lipocalin as an early biomarker for canine renal ischemia-reperfusion injury.

BackgroundThe pathological mechanism of ischemia/reperfusion acute kidney injury (I/R-AKI) differs from other forms of AKI. Neutrophil gelatinase-associated lipocalin (NGAL) is a sensitive biomarker for early diagnosis of AKI, but its utility for diagnosis of canine I/R-AKI remains to be evaluated. The aims of this study were to establish an I/R-AKI model in dogs and to evaluate the diagnostic value of NGAL for canine I/R-AKI.MethodsWe randomly divided 12 beagle dogs into a sham and an I/R group. Artery and vein of the left kidneys of I/R group were cross-clamped for 60 min followed by reperfusion. The kidney samples were analyzed for histopathological lesions. Serum and urinary samples were analyzed for blood urea nitrogen (BUN), serum creatinine (sCr), serum NGAL (sNGAL), urinary creatinine (uCr), and urinary NGAL (uNGAL). Their detection sensitivities and specificities were compared using a receiver operating characteristics (ROC) method. The expression of NGAL in the renal tissues was analyzed by quantitative RT-PCR (qRT-PCR) and immunohistochemical (IHC) analysis.ResultsAfter I/R, histopathological analysis showed typical AKI lesions in the dog kidneys of the I/R group, but not in the sham group. Compared to that of the sham group, BUN and sCr of the I/R group rose to significant high levels from 24 h after I/R. Both uNGAL and sNGAL rose rapidly from 2 h, reached to the peak levels at 12 h, and then receded to the pre-operation levels by 72 h after I/R. The uNGAL/uCr ratio (uNCR) rose rapidly from 2 h and remained at variably high levels from 6 to 60 h after I/R. The ROC analysis showed that detection sensitivities of uNCR, uNGAL, and sNGAL were significantly (P<0.0001) higher than that of sCr, without significant difference in specificity. The cut-off values of sNGAL, uNGAL and uNCR were 14,642 pg/mL, 6,773 pg/mL, and 6,701 pg/mg, respectively. Both qRT-PCR and IHC analyses confirmed the dynamic expression of NGAL in the dog kidneys with ischemic acute kidney injury (I-AKI).ConclusionsThere is potential for NGAL to be used as a sensitive biomarker for early diagnosis of canine I-AKI.

Identification of key metabolites during cisplatin-induced acute kidney injury using an HPLC-TOF/MS-based non-targeted urine and kidney metabolomics approach in rats

Kidney injury is a major adverse effect of cisplatin use. Metabolomics has been used to characterize physiological or pathological conditions through identification of metabolites and characterization of the metabolic pathway. Metabolomics profiling could allow for identification of nephrotoxic mechanisms of cisplatin and identification of biomarkers of cisplatin-induced injury. In this study, we performed metabolomics analysis to characterize key changes in metabolite levels during cisplatin-induced acute kidney injury (AKI) in rats, and screened for sensitive biomarkers for early diagnosis using HPLC-TOF/MS. Rats were intraperitoneally injected with 7.5 mg/kg or 15 mg/kg of cisplatin, or normal saline, and 12 h urine and kidney samples were collected after 72 h. Serum biochemical parameters and kidney histological evaluations showed dose-dependent AKI in response to cisplatin. Metabolomics analysis showed that 37 and 35 endogenous metabolite levels changed in rat urine and kidneys, respectively. Seven key metabolic pathways were disrupted, including the tricarboxylic acid cycle (TCA cycle), phenylalanine, tyrosine, and tryptophan biosynthesis, phenylalanine metabolism, glycerophospholipid metabolism, taurine and hypotaurine metabolism, d-glutamine and d-glutamate metabolism, and nicotinate and nicotinamide metabolism. These pathways are involved in energy generation, and amino acid and lipid metabolism, and disruption of these pathways could contribute to oxidative stress injury, inflammation, and cell membrane damage. Furthermore, 11 sensitive metabolites in urine were screened as potential biomarkers of AKI. To validate these biomarkers, we quantified 4 off these biomarkers, and confirmed that levels of these metabolites were altered in urine of rats treated with CDDP.

Comprehensive Transcriptome Profiling of Peripheral Blood Mononuclear Cells from Patients with Sepsis

Background: Sepsis, as a clinical emergency, usually causes multiorgan dysfunction and can lead to high mortality. Establishment of specific and sensitive biomarkers for early diagnosis is critical to identify patients who would benefit from targeted therapy. In this study, we investigated this syndrome by analyzing the transcriptome of peripheral blood mononuclear cells (PBMCs) from patients with sepsis and identified sepsis-specific biomarkers.Methods: In this study, a total of 87 patients with sepsis and 40 healthy controls from a prospective multicenter cohort were enrolled. Samples from 44 subjects (24 patients with sepsis and 20 healthy controls) were sequenced and the remaining patients were included in the validation group. Using high-throughput sequencing, a gene expression profile of PBMCs from patients with sepsis was generated to elucidate the pathophysiology of sepsis and identify sepsis-specific biomarkers.Results: Principal component analysis (PCA) and unsupervised hierarchical cluster analysis showed that patients with sepsis separated from healthy controls. A total of 1639 differentially expressed genes (DEGs) were identified (|log2 fold change|>2, adjusted P value <0.05) between these two groups, with 1278 (78.0%) upregulated and 361 (22.0%) downregulated in patients with sepsis. Gene Ontology (GO) analysis of the upregulated DEGs identified 194 GO terms that were clustered into 27 groups, and analysis of the downregulated DEGs identified 20 GO terms that were clustered into 4 groups. Four unique genes were identified that could be predictive of patients with sepsis. External validation of the four genes using quantitative real-time polymerase chain reaction (qRT-PCR) was consistent with the results of mRNA sequencing, revealing their potential in sepsis diagnosis.Conclusions: The transcriptome characteristics of PBMCs, which were significantly altered in sepsis patients, provide new insights into sepsis pathogenesis. The four identified gene expression changes differentiated patients with sepsis from healthy subjects, which could serve as a convenient tool contributing to sepsis diagnosis.

Lnc-TCL6 is a potential biomarker for early diagnosis and grade in liver-cirrhosis patients.

BackgroundLong non-coding RNAs (lncRNAs) have been applied as biomarkers in many diseases. However, scarce biomarkers are available in single lncRNA differential expression associated with different clinical stages of liver cirrhosis (LC). The aim of the study is to identify some lncRNAs that can serve as non-invasive sensitive biomarkers for early diagnosis and grade of LC.MethodsBlood lncRNA expression was evaluated in three independent cohorts with 305 participants including healthy controls, hepatitis B virus (HBV) carriers, and patients with chronic hepatitis B (CHB) or LC. First, candidate lncRNAs were screened by CapitalBiotech microarray to diagnose cirrhosis. Quantitative reverse-transcriptase polymerase chain reaction was then used to investigate the expression of selected lncRNAs in the whole group of cirrhosis and different Child–Pugh classes. Ultimately, the diagnostic accuracy of the promising biomarker was examined and validated via Mann–Whitney test and receiver-operating characteristics analysis.ResultsLnc-TCL6 was identified as a sensitive biomarker for early diagnosis of LC (Child–Pugh A) compared with healthy controls (area under the ROC curve [AUC] = 0.636), HBV carriers (AUC = 0.671), and CHB patients (AUC = 0.672). Furthermore, lnc-TCL6 showed a favourable capacity in discriminating among different Child–Pugh classes (AUC: 0.711–0.837). Compared with healthy controls, HBV carriers, and CHB patients, the expression of lnc-TCL6 was obviously up-regulated in Child–Pugh A patients and, conversely, significantly down-regulated in Child–Pugh C patients.ConclusionsLnc-TCL6 is a novel potential biomarker for early diagnosis of LC and is a possible predictor of disease progression.

The levels of urine CTX-II, C2C, and PYD in children patients with Kashin-Beck disease in Qinghai Province of China

BackgroundKashin-Beck disease (KBD) is an endemic and chronic osteoarthropathy. At present, the diagnosis of KBD mainly depends on the X-ray examination and which could not reflect early damage of cartilage sensitively. So, the aim of this study was to find effective and sensitive biomarkers for early diagnosis of pediatric KBD.MethodsA total of 122 children aged 7–15 years old from 3 villages of Qinghai Province were eligible for the study. Thirty-one, 41, and 50 children were assigned in case, internal, and external control groups, respectively. The levels of CTX-II, C2C, and PYD in urine were measured by using ELISA and compared statistically. In addition, the receiver operating characteristic curve (ROC) analysis was used to assess the performance of diagnostic biomarkers.ResultsThere were significant differences in levels of CTX-II, C2C, and PYD in urine of subjects among three groups. The levels of CTX-II and PYD in the case group were significantly higher than those in external and internal control groups. On the contrary, the level of C2C in the case group was lower than that in the external control group. Compared to the external control group, the area under the curve (AUC) of CTX-II, C2C, and PYD were 0.857, 0.837, and 0.79, and the AUC of CTX-II significantly higher than that of PYD. Compared to the internal control group, the AUC of CTX-II, C2C, and PYD were 0.911, 0.875, and 0.839, and there were no significant differences in the AUC among three indicators.ConclusionBoth CTX-II and PYD in urine could be used as biomarkers for early diagnosis of pediatric KBD, and the prediction accuracy of CTX-II was relatively superior.

The role of biological oncomarkers in the differential diagnosis of complex kidney cysts

The renal cell carcinoma (RCC) ranks first place in mortality among urogenital tumors, and the incidence is third after prostate and bladder cancer. The cystic form of renal cell carcinoma is 5–7%, and according to recent data – even 10% of all kidney tumors. The early diagnosis of RCC allows for effective treatment, which significantly increases survival.Early and accurate diagnosis helps to avoid inadequate treatment, provides a favorable prognosis for progression of the disease and allows for more effective therapy. Most of the renal tumors, including cystic, are diagnosed accidentally during an examination for other reasons. Small tumors are usually asymptomatic, which leads to late diagnosis, and consequently, to the low effectiveness of treatment.Thus, the need for the use of sensitive biomarkers for early diagnosis of RCC and monitoring of its progression is clearly followed. Nowadays, many attempts have been made by scientists to identify a new informative kidney tumor biomarker that could be used for early diagnosis and disease progression, as well as prognostic capabilities. This overview summarizes the latest advances in the discovery of these biomarkers and their clinical value.

Urinary RKIP/p-RKIP is a potential diagnostic and prognostic marker of clear cell renal cell carcinoma.

Clear cell Renal Cell Carcinoma (ccRCC) causes over 13,000 deaths each year, and about 20,000 new cases/year in Europe. In most cases, the causes are unknown and, most importantly, there are no reliable biomarkers for the diagnosis and prognosis of this disease. The search for sensitive biomarkers for early diagnosis and prognosis of clear cell Renal Cell Carcinoma (ccRCC) is currently a fast growing field. We carried out proteomics analysis of 93 urinary samples of healthy subjects (HS) and patients affected by ccRCC, prostate cancer (PCa) and chronic kidney disease (CKD), that was able to successfully distinguish each group.The most significant candidate biomarker was identified by mass spectrometry as Raf Kinase Inhibitor Protein (RKIP), a key regulator of cell signaling, already described in several cancer types as a metastasis suppressor. By combining ELISA, immunoblotting and tissue microarray, we demonstrated that, in ccRCC, urinary excretion of RKIP and its phosphorylated form (p-RKIP) reflected the tissue expression of these putative biomarkers. Baseline urinary RKIP, evaluated in an independent cohort of 56 ccRCC patients and 28 HS, successfully distinguished both groups and, most importantly, a cut-off value of 10 ng/mg/g Pr/uCr enabled a highly accurate prediction of Cancer-specific survival and Progression-free survival. Furthermore, p-RKIP was totally undetectable in both tissue and urine samples of ccRCC, showing a great potential for diagnostics purposes.Our data indicate that urinary RKIP encompasses both the unphosphorylated and the phosphorylated form and that their combined evaluation can help in the diagnosis and prognosis of ccRCC.

EyeGuidance – a computer controlled system to guide eye movements

Abstract The densely innervated human cornea is the only superficial tissue of the human body in which nerve fibres are accessible in vivo by corneal confocal microscopy (CCM). Morphological parameters of the corneal sub-basal nerve plexus (SNP) derived from CCM images can potentially serve as a sensitive biomarker for early diagnosis of various neurodegenerative diseases. The evaluation of a single image with a typical field of view of 0.16 mm2 is insufficient for robust morphometric assessment. Mosaicking approaches have therefore been proposed to examine the SNP on a larger scale. Here we present a highly automated technique that significantly facilitates the generation of mosaic images of the SNP and is suitable for clinical tests.

A study of carcinoembryonic antigen concentrations in patients with coronary artery disease

Abstract Background & aims Carcinoembryonic antigen (CEA) is a tumour marker whose role is now being evaluated as a biomarker for early diagnosis of acute coronary syndromes. Its levels may rise even prior to rise of established biomarkers of myocardial necrosis. Methods Serum CEA concentrations were measured by double sandwich ELISA method in four groups of subjects with STEMI, NSTEMI/UA (24 patients of NSTEMI and 18 patients of unstable angina), stable angina and healthy controls with 42 males between 40 to 60 years of age in each group. Also qualitative Troponin T assay and CPK-MB concentrations measurement was done in groups with STEMI and NSTEMI/UA and correlations between CEA concentrations and Troponin T and CPK-MB were made. Results Mean serum CEA concentrations were 5.19 ± 0.39, 3.84 ± 0.34, 3.91 ± 1.40, 1.84 ± 0.35 and 2.12 ± 0.40 ng/ml respectively in groups with STEMI, NSTEMI/UA, UA alone, stable CAD and healthy controls respectively. STEMI, NSTEMI/UA and UA patients had significantly higher concentrations than stable CAD and healthy controls (p Conclusions CEA is a sensitive biomarker for early diagnosis of ACS. Its levels correlate with severity of ACS. It may play a crucial role in diagnosis of ACS in patients who present with atypical manifestations or when traditional biomarkers such as CPK-MB and Troponins are not raised.

Cadmium-induced genomic instability in Arabidopsis: Molecular toxicological biomarkers for early diagnosis of cadmium stress

Microsatellite instability (MSI) analysis, random-amplified polymorphic DNA (RAPD), and methylation-sensitive arbitrarily primed PCR (MSAP-PCR) are methods to evaluate the toxicity of environmental pollutants in stress-treated plants and human cancer cells. Here, we evaluate these techniques to screen for genetic and epigenetic alterations of Arabidopsis plantlets exposed to 0–5.0 mg L−1 cadmium (Cd) for 15 d. There was a substantial increase in RAPD polymorphism of 24.5, and in genomic methylation polymorphism of 30.5–34.5 at CpG and of 14.5–20 at CHG sites under Cd stress of 5.0 mg L−1 by RAPD and of 0.25–5.0 mg L−1 by MSAP-PCR, respectively. However, only a tiny increase of 1.5 loci by RAPD occurred under Cd stress of 4.0 mg L−1, and an additional high dose (8.0 mg L−1) resulted in one repeat by MSI analysis. MSAP-PCR detected the most significant epigenetic modifications in plantlets exposed to Cd stress, and the patterns of hypermethylation and polymorphisms were consistent with inverted U-shaped dose responses. The presence of genomic methylation polymorphism in Cd-treated seedlings, prior to the onset of RAPD polymorphism, MSI and obvious growth effects, suggests that these altered DNA methylation loci are the most sensitive biomarkers for early diagnosis and risk assessment of genotoxic effects of Cd pollution in ecotoxicology.

Role of biomarkers of nephrotoxic acute kidney injury in deliberate poisoning and envenomation in less developed countries.

Acute kidney injury (AKI) has diverse causes and is associated with increased mortality and morbidity. In less developed countries (LDC), nephrotoxic AKI (ToxAKI) is common and mainly due to deliberate ingestion of nephrotoxic pesticides, toxic plants or to snake envenomation. ToxAKI shares some pathophysiological pathways with the much more intensively studied ischaemic AKI, but in contrast to ischaemic AKI, most victims are young, previously healthy adults. Diagnosis of AKI is currently based on a rise in serum creatinine. However this may delay diagnosis because of the kinetics of creatinine. Baseline creatinine values are also rarely available in LDC. Novel renal injury biomarkers offer a way forward because they usually increase more rapidly in AKI and are normally regarded as absent or very low in concentration, thereby reducing the need for a baseline estimate. This should increase sensitivity and speed of diagnosis. Specificity should also be increased for urine biomarkers since many originate from the renal tubular epithelium. Earlier diagnosis of ToxAKI should allow earlier initiation of appropriate therapy. However, translation of novel biomarkers of ToxAKI into clinical practice requires better understanding of non-renal factors in poisoning that alter biomarkers and the influence of dose of nephrotoxin on biomarker performance. Further issues are establishing LDC population-based normal ranges and assessing sampling and analytical parameters for low resource settings. The potential role of renal biomarkers in exploring ToxAKI aetiologies for chronic kidney disease of unknown origin (CKDu) is a high research priority in LDC. Therefore, developing more sensitive biomarkers for early diagnosis of nephrotoxicity is a critical step to making progress against AKI and CKDu in the developing world.

Protein Map Standardization of Human Saliva Using Two Dimensional Gel Electrophoresis (2-DE)

Over the past decade, the use of saliva as an auxiliary diagnostic tool for biomarker detection has gained considerable acceptance as a non-invasive, inexpensive alternative to conventional serum method. In proteomics, the most promising technique which can be used for detecting salivary biomarker with sufficient resolving power is two-dimensional gel electrophoresis (2-DE) that provides a unique platform for the simultaneous separation of proteins in a complex mixture. However, as a fact most of the new scientists in developing countries are still facing problems with optimization of 2-DE techniques because the performance of optimization techniques in proteomics research using 2-DE has its own limitations. Therefore, the present study was established to generate a reproducible and optimized protocol to display the 2-DE protein map of human saliva. Our results showed the standard optimization of the 2-DE protein mapping was achieved at pH 3-10 with 60 μg proteins loading. This protocol could be used by other scientists along with identification of differentially expressed proteins by mass spectrometry when 2-DE protein map of patients saliva are compared to that of normal healthy individuals. These differentially expressed proteins could be later used as specific and sensitive biomarkers for early diagnosis and prognosis of the disease in question. In conclusion, the procedure used in our study generated a highly reproducible and optimized reference 2-DE protein mapping of human saliva.